Discovery and characterization of lung cancer biomarkers using monoclonal antibody proteomics and liquid chromatography: mass spectrometry of glycopeptides.

• Online Access: http://hdl.handle.net/2047/d20000801

Current proteomics strategies for biomarker efforts have two limitations: the neglect of protein glycosylation in discovery/qualification and the lack of antibodies for verification/validation. The main goal of this thesis is to address these two problems. For the lack of high quality antibodies, monoclonal antibody (mAb) proteomics - high throughput mAb generation and disease -specific mAb screening - was applied to lung cancer biomarker discovery. mAb proteomics is demonstrated to generate high quality mAbs in a high throughput manner, an important characteristic for biomarker verification/validation. For glycosylation characterization, mass spectrometry (MS)-based methods for sensitive, site-specific discovery and quantitation were developed and applied to characterize the glycosylation changes of the lung cancer biomarker candidate haptoglobin. These developed techniques for glycosylation characterization can be also applied to other glycoproteins and therefore will contribute to the emerging glyco biomarker field.